Using a method of retroviral gene transduction we have been able to generate conditionally immortalized long-term hematopoietic stem cell lines from primary human cord blood stem cells. Phase I of this project includes applying newly discovered non- viral technology to generate human cell lines and the subsequent differentiation of those cells into human red blood cells. In addition, the project includes testing the function of these cells in small animals. Phase II of the project will entail optimizing large-scale production of protein transduced long-term hematopoietic stem cells and large-scale production of red blood cells for eventual testing in larger animals and human clinical trials. PUBLIC HEALTH RELEVANCE: This project seeks to optimize a novel method to generate large amounts of human red blood cells for clinical use from human adult stem cells. Once this method has been validated for large-scale production, the long-term goal for benefiting public health will be to use this system to generate red blood cells for clinical applications across a variety of global health conditions including, but not limited to, blood disorders, surgeries and in cases of acute trauma.